21-diazo pregnenes



United Stotes Patent 2,904,545 '21- DIAZO ,PREGNENES TatleusfReichstein,Albert Wettstein, Georg Aimer, Jean- Ren Biileter, {and ,Karl Heusler,Basel, Robert Neher, Binningen, .Julii s v'Schmidlin, Jlasel, ;Hel l n tUeberwasser, Riehen, and ,Peter .Wielaml, Basel, Switzei land; saidWettstein, said .A me ,said ,Billeter, said fieg sler, saidfNehesaildjschnlidlin, sai l' U eberwassei and said Wieland, assignors tosai'd'Tarleus Reichstein No Drawing. Application January 5,1955

Serial-No. 480 061 Claims priority, t pplicationswitze 'land Janua y15,1254 8 Claims. (Cl. 260--'g23 9.55)

Serial No. 444,65'7,'filed July 20, 1954, by T.:Reichstein Aldoste onelNaJo. or

Pgtented Sept. 15, 19 59 i t zni v l imv s A1dosterone monoacetate (morereadily esterifiable hydroxyl group is acylated) lCrO;

T W ltf -Kjshner lNaIO;

0o coon COOH J10 H3O a no i 1. CHzNq 2. Hz, Pt 3. 010

00 .coocn, oooolzt Instead of aldosterone, there may be used as startingmaterial functional derivatives thereof in which both hydroxyl groupsare free or in which one hydroxyl group is functionally converted,preferably esters, or ethers, enol esters, enol ethers, acetals, ortheir corresponding thio derivatives, such as thio ethers and thioacetals.

By the degradation reactions shown in the above diagram of formulae,aldosterone has the following formulae which obviously represent anequilibrium l H H nron OH ,0 ono These formulae show that aldosteroneand its functional derivatives are 3,1l-dioxygenated pregnane compoundsor functional derivatives thereof, 'in which the radical in l3-positionis a free or functionally converted aldehyde group. Furthermore, it waspossible to prove the constitution of aldosterone by the process for themanufacture of such compounds as described below.

Steroids of this kind are obtained when in 3-oxygenated, specially3,11-dioxygenated steroid-l7-carboxylic acids, or their functionalderivatives, in which the radical in 13- position is a free orfunctionally converted aldehyde group or a radical convertiblethereinto, the radical in the 17- position is converted into the acetylgroup or into the free or a functionally converted hydroxy acetyl group;e.g. halides of the acids mentioned are reacted with diazo methane; ormethyl metal compounds are caused to act on functional derivatives, suchas halides, esters, acid amides or nitriles; a free or functionallyconverted hydroxyl group is introduced into the 21-position of thecompounds obtained or a substituent present in the 21-position isconverted into such group, or any substituent in 21- position isreplaced by hydrogen, or a methylene group ing at carbon atom 20 isoxidized into an oxo group, if desired, a free or functionally convertedhydroxyl group is introduced into the l7-position, if there is no freeor functionally converted aldehyde group in 13-position, no oxo group in3-position, and no double bond in 4,5-position, these are formed, ifdesired, at any stage of the process, and free or functionally convertedhydroxyl or oxo groups are converted into one another.

In carrying out the reactions of this process per se conventionalmethods may be followed. Thus the reaction of the acid halides withdiazo methane can be carried out with an excess of the latter to obtainthe diazo ketones, hydrogen halide being split off. On the other hand,when the diazo methane solution is gradually added to the acid halide,the hydrohalic acid liberated during the condensation reacts with theintermediately formed diazo ketone, and the corresponding halogen ketoneis obtained.

For the replacement of the halogen atoms or the diazo group in theresulting ketones by hydrogen a large number of reducing agents aresuitable, for example, metals, such as zinc, copper, magnesium or theiralloys in the presence of any of a wide variety of solvents, such asalcohols, aqueous alcohols, lower aliphatic acids, hydrohalic acids, oralkalis. Reductions of this kind are often carried out electrolyticallyor with hydrogen catalytically activated, e.g. with nickel, palladium orplatinum. Finally, it is also possible to effect the reduction withaluminium amalgam or sodium amalgam, zinc palladium, alkali iodides andglacial acetic acid, sodium and alcohols, disubstituted anilines or withcombinations of the above reducing agents. The reduction proceedsespecially easily in the case of the'2l-iodinated products.

To form hydroxy ketones, diazo ketones obtained are treated, either inthe crude state or after separation and purification, with hydrolyzingagents, for example, water or dilute acids, as e.g. sulfuric acid, orwith organic sulfonic acids, such as methane sulfonic acid or toluenesulfonic acid. Esters of the hydroxy ketones can be obtained by reactingthe diazo ketones with organic or inorganic acids having a low watercontent, for example, acetic acid, or with propionic acid, butyric acid,trimethyl acetic acid, crotonic acid, oenanthic acid, palmitic acid,benzoic acid, phenyl acetic acid, ,B-cyclo-pentyl propionic acid,hydrochloric acid, hydrobromic acid, hydriodic acid, phosphoric acid, orboric acid. When halogen ketones are obtained as intermediate products,they can be converted by means of alkaline agents, for examplebicarbonates, into the free hydroxy ketones or by means of salts of theaforementioned acids into their esters.

The hydroxyketones and their esters can also be obtained from the methylketones either directly or indirectly. Thus, they can be obtained withthe aid of suitable oxidizing agents, for example lead tetracylates,such as lead tetracetate or aryliodoso acylates. It is also possible tohalogenate the methyl ketones in 21- position directly, or indirectly byway of the 21-oxalyl derivatives, and converting the resulting halogenketones into the hydroxy ketones or their esters, following the abovedirections.

According to this process, the acid halides mentioned can also bereacted with methyl metal compounds, for example those of magnesium,cadmium or zinc; there may also be mentioned compounds of mercury,copper, aluminium, tin and the alkali metals. There are thus .obtainedthe methyl ketones which can be converted into the hydroxy methylketones in the afore-described manner.

When acid esters are converted with the methyl metal compounds, thereare obtained, with the splitting oil? of water, from the intermediatelyformed carbinols compounds having a methylene grouping at carbon atom20. The latter grouping can be replaced by an oxo group by means ofoxidizing agents. There may be mentioned by way of example the oxidationwith a compound of hexavalent chromium, such as chromic acid, withpermanganate, by ozonizing and splitting the ozonide, by the action ofperoxides, such as perbenzoic acid, monoperphthalic acid, or hydrogenperoxide, advantageously in the presence of osmium tetroxide and thesplitting, for example by means of lead tetracetate or periodic acid, ofthe glycols formed in the hydrolysis of the oxide rings or by directadditive combination of 2 hydroxyl groups with the double bond.

In the reaction of the nitriles with methyl metal compounds iminocompounds are obtained which can be hydrolyzed into ketones withhydrolyzing agents.

If desired, the free or functionally converted hydroxyl group can beintroduced into the l7-position of resulting pregnane compounds by knownmethods at any stage of the process.

When the 13-position of the compound obtained is occupied by a radicalwhich is convertible into a free or functionally converted aldehydegroup, such as a free or functionally converted carboxyl group,especially a lactone group, or a free or functionally converted carbinolgroup, an agent suitable for this kind may be employed, such as e.g. areducing or oxidizing agent. Depending on the reaction conditions18,11-lactones, for example, are converted by means of alkali aluminiumhydrides, such as lithium-aluminium hydride into 11- hydroxy-aldehydesor their cyclosemiacetals or 11,18- dioles.

According to this invention, a 3-, 11- and/ or l8-positioned substituentconvertible by hydrolysis into an oxo or hydroxyl groupv can be soconverted at any stage of the process. For example, ketals and acetals,open chain as well as cyclic ones, e.g. ethylene ketals, can be split bytreatment with mineral acids or sulfonic acids at 5 i oom temperature,advantageously in the presence of a ketone; such as acetone orv pyruvicacid, or by heating gently with dilute acetic acid; Under the same acidconditions, enol ethers or tetrahydropyranyl ethers are also split.Benzyl ethers can moreover be split easily with hydrogen in the presenceofa catalyst, e.g. palladium on carrier substances, such as animalcarbon or alkaline earth carbonates.

The conversion of the free hydroxyl groups into 'oxo groups is effected,e.g. by means of oxidizing agents, such as chromic acid in glacialacetic acid, or by dehydrogenating means, such as heatingwith copperpowder, the action of metal alcoholates or metal phenolates in thepresence of ketones, such as acetone or cyclohexanone.

Hydroxyl or oxo groups may also be functionally converted as described,eg in the aforementioned Serial No. 454,834, filed September 8, 1954, byT. Reichstein et a1.

If the double bond must be introduced into the 4,5- position, this canbe done in the usual manner, for example by halogenating andsubsequently splitting off hydrogen halide.

Racemates obtained can be split into their antipodes by known methods atany stage of the process.

The compounds obtained by the process of this invention are saturated orunsaturated I i-oxygenated, especially 3,11-dixygenated pregnenederivatives, of which the radical in 13-position is a free orfunctionally converted aldehyde group or a residue convertiblethereintol' Oxygenated compounds and their functional derivatives areunderstood to be such as contain free or functionally converted hydroxylor 0x0 groups, such as eg esters, ethers, thio esters, thio ethers,thiol esters, thione esters, acetals, mercaptals, ketals, enolderivatives, such as enol esters, enol others or enamines, hydrazones,semicarbazones and the like.

The products of the present process may be used as medicaments or asintermediate products for making medicaments.

The present invention also relates to any modification Startingmaterials for the process of this invention are acid derivatives, suchas halides, esters, acid amides, nitriles of 3-oxygena ted, especially3,1l-di0xygenated steroid-l7-carboxylic acids, including compounds ofthe homo and nor series, especially l9-nor and/ or D-homo- :acetianicacids, the carboxyl group of the latter being in the 170t-POSitiOI1, ofcourse. of any steric configuration and contain at the carbon atom 3 andespecially carbon atoms 3 and-11 free or functionally converted hydroxylor oxo groups and in 13-position a free or functionally convertedaldehyde group or a residue convertible thereinto, such as for example afree or functionally converted carboxyl or carbinol group. Thus theresidue in l3-position may for example be a lactonized carboxyl group,e.g. lactonized with the llB-hydroxyl. The starting materials aresaturated and may contain a double bond extending, for example, fromcarbon atom 5, in other words, beingin the 4,5 or 5,6-position.

The starting materials are new. They can be made for example by theprocess of our application-Serial No. 480,062, filed concurrentlyherewith. Moreover, some of them are obtained whenisaturated orunsaturated 3,11- dioxygenated 17- or 20-keto-steroids or 3-oxygenated13,17-seco-steroid-l7,l3-lactones are oxidatively converted into3,1l-dioxygenated 13,17-seco-steroid-l7,13- lactones, the latter cleavedinto l3-carbinols, water is eliminated directly orindirectly, if desiredafter the preparation of an ll-keto group, the side chain in 0a,,B-UI1-The starting materials are saturated ketones obtained is degraded,ifdesired, iii known manner to 2'or 3 carbon atoms and/or converted intoa substitue'nt'whichreacts with "anace'tic 'acid'deriva-' tivecontaining an active methylene 'group, the methyl group at thedouble'bond is oxygenated and the com pounds obtained are reacted toform 3,11,l'8-"trioxy= genated' steroid-17-carboxylic' acids orpregnant:'-20-on"es or functional derivatives. byway 'dfsubstitut'edacetic' acid derivatives having "an 'activernethinegroup.

These starting materialsfare oxidatively 'convefieddnto thecorresponding 3;11-droxygenated43:17-see steroid: 17 :13-lactones, forexample; by means of hydropero'x ides, such as hydrogen peroxide orpe'ra'cids, for exain ple, p'erformic acid, per'aceticacid o'r"perbenzoic ac or by a microbiologicalrnethod; for 'example, by means ofmicro-organisms of the group's Mucorales, such as Cunninghamella,Actinomyce'te's, such asstr mmyeetes; 'Curvularia, Penicillium,Cylindrocarpb'n, 'Gliocladium', Aspergillus types or Fungi irnperf'ecti.Thelatt'ef method is also especially suitable for theintioduction'o'f'oxygen into the ll-pos'ition of lactones'to be u'sedintheproc'ess ofthe invention or for the degradation of Z'O 'ke'ton'eS to lactones.During these and thesubsequent 'oxidations, especially whenchemical'oxidiz'ing' agents are used," re active double bonds inunsaturated rings 'of'th'e starting materials, for example, a doublebond in th'e'5'z'6-positioii', are protected, if desired,intermediately, for example, by additive combination with halogen or ahydrogen halide or conversion into an i-steroid. Any free hydroxylgroups present are advantageously temporarily protected wholly or inpart, for example, by esterification or etheri- I fication.

A further step in the process consists in splitting up the lactone soobtained into a 13-carbinol, especially by means of a hydrolyzing agentsuch as an alkali hydroxide, an alkaline earth metal hydroxide,aluminium oxide and the like, or with an organo-metal compound such as aGrignard compound, for example methyl-, or phenylmagnesium halide, or analkylor aryl-lithium compound. If the treatment is carried out with onlyabout one equivalent of alkyl magnesium halide, it is at first possibleonly to cause the addition of an alkyl residue without splitting up thering. A

After a different substituentin the ll-position has been converted inknown manner into a keto group, the elimination of the 13-hydroxyl grouptakes place very easily. Normally it takes place directly withelimination of water, generally with the reagents used for splitting upthe lactone ring. In thecase of the dihydropyrane derivatives or thecorresponding 7-ring compounds which are at first obtained, if desired,with alkyl magnesium halides, water is in effect split off indirectly bythe oxidation and splitting otf'of the acyloxy group in the 13-position.After the splitting up of the lactone ring by means of at least twoequivalents of organometal compound, and simultaneously with thesplitting off of water in the nucleus or independently thereof, watermay be split off in the side chain, for example, by means of an excessof the organo metal compound or by an agent customarily used forsplitting off water.

A further step which may be carried out if desired, is the conversion ofa 16-substituent'in'to a hydroxyl, oxo or amino group, especially bymeans of a hydrolysing agent. The side chain in-tlie mil-unsaturatedketone is then degraded in a manner in itself known to two carbon atoms,or if D-homo-compounds are desired to three carbon atoms, and/0r.converted into a substituent which reacts with an acetic acid derivativecontaining an active methylene group. For the degradation there isadvantageously used an oxidising agent such as ozone, chromic acid orlead tetracetate, or the silver saltof the carboxylic acid may bedegraded by means of halogen by themethod of Hunsdiecker.

In order to oxygenate the methyl group at the double bond it may, forexample, be hydroxylated or acyloxylated, directly, for example, bymeans of selenium dioxide, lead tetracetate, or indirectly,.for example,by halogenation, ketone splitting and subsequent reaction, for example,by means of a salt of a carboxylic acid such as an alkali acylate orsilver acylate. If it is desired, for example, to bring aboutoxygenation, this step may be carried out by oxidation of thehydroxylated compound or directly. Thus, for example, the startingcompound may be reacted with selenium dioxide or the above mentionedhalide may be converted by the method of Krohnke by way of thequaternary ammonium salt and the nitrone. In this case also derivativesof the aldehyde group may be obtained intennediately, for example,acetals with monohydric or polyhydric alcohols, and also diacylates. Theaforesaid total conversion of the methyl group at the double bond mayalso be carried out at a different stage in the synthesis and may beinterrupted by other later process steps.

Likewise, at any intermediate stage in the synthesis, a A -3-keto groupmay be introduced in known manner, especially by halogenation and thesplitting off of hydrogen halide, for example, by means of a hydrazinesuch as semicarbazide or by means of pyruvic acid, and the oxidation ofa 3-hydroxyl group and the halogenation may be carried out in a singlereaction by means of an alkyl hypohalide. The A -3-keto group soproduced may,

if desired, be subsequently protected intermediately, for example, byketalisation or the formation of an enol derivative by a method initself known.

An essential step of the process is the conversion to a steroid by wayof a substituted acetic acid derivative containing an active methinegroup, such as an aceto: acetic acid derivative or malonic acidderivative. This is carried out, for example, by subjecting a primary16- halide or pseudohalide to a reaction of' the character of anacetoacetic acid ester or malonic acid ester synthesis and tointramolecular condensation according to the method of Michael, or byfirst carrying out a Michael condensation and subjecting the resultingacetic acid de rivative to ring closure. As is usual in acetoacetic acidester and malonic acid ester syntheses the resulting derivativecontaining one or two substituted acid residues is subjected in knownmanner to a ketone or acid splitting.

In a stage following ring closure to form the steroid the ll-keto groupcan reductively be converted into an ll-hydroxyl group, advantageouslyan 11,8-hydroxyl group, for example, by means of a light metal hydridesuch as an alkali metal boron hydride or alkali metal aluminium hydride.

One form of the process for the preparation of the starting materials isillustrated by the following diagram of formulae /Ph x Ph \fi A00 OHPh-MgBr H0 AcO- A004 EO- I x=n,o1=r II III AcOH, ACzOJy I Ph oooom ooonV= CHzNg oro I P11 Ac0 AeO- A00- vr V W lNBS 0mm- 0mm GH OAc o 0- 0oooon, oooon, oooorn V orr orr, HBr KOAc in acetone AcO- HO- no-- VIIVIII 1x 1 l The resulting compound XXV, on ketal and acetal splittingand subsequent oxidation yields the 18,11-1actone of A-3-oxo-1lfi-hydroxy-etienic acid-18-acid obtained by degradingaldosterone The following examples illustrate the invention:

Example 1 A solution of 0.531 gram of dry A-3zl8-bis-ethylenedioxy-llfi-trifiuoracetoxy-aetienic acid in 15 cc. ofabsolute benzene and 0.8 cc. of absolute pyridine is mixed at 5 C. with1.8 cc. of oxalyl chloride in cc. of absolute benzene, and the whole isallowed to stand at 20 C. for 20 minutes. It is then evaporated invacuo, the residue is taken up in 20 cc. of absolute benzene, and themixture is filtered to remove the insoluble precipitate. The filtrate isintroduced into 26.5 cc. of ice-cold dry 0.38-molar ethereal solution ofdiazo-methane, and then allowed to stand at room temperature for 2hours, and evaporated in vacuo. There is obtained crude A 3:18 bisethylene dioxy 11/3 trifluoracetoxy 20 keto- 21-diazopregnene.

The diazo-ketone so prepared is dissolved in 30 cc. of methanol, thenmixed with 0.800 gram of potassium carbonate in 12 cc. of water, andallowed to stand in the molten state in vacuo for 22 hours at roomtemperature. It is then diluted with water, the methanol is removed invacuo, and the suspension remaining behind is extracted by agitationwith a mixture of ether and chloroform, (3:1). The solution is washedwith water and dried with sodium sulfate, and then evaporated in vacuoto yield crude A-3z18-bis-ethylene-dioxy-11,8-hydroxy-20-keto-2l-diazo-pregnene.

The resulting free llfi-hydroxy-compound is heated with 4 cc. of glacialacetic acid for 30 minutes at 100 C., after which the splitting off ofnitrogen ceases. The mixture is then evaporated in vacuo, the residue isdissolved in 120 cc. of acetone, 0.076 gram of para-toluene sulfonicacid is added, and the mixture is stirred for 24 hours at roomtemperature in an atmosphere of nitrogen. The reaction mixture is cooledin ice water and mixed with 4.0 cc. of an 0.1 N-solution of sodiumcarbonate and 30 cc. of water, and the acetone is distilled off invacuo. The suspension which remains behind is extracted by agitatio-nwith a mixture of ether and chloroform (3:1), the extract is washed withWater, dried with sodium sulfate and evaporated in vacuo. The residue isdissolved in benzene, and the solution is chromatographed over a columnof grams of aluminium oxide prepared with benzene. There are used aselutriating agents benzene, a mixture of benzene and ether (3:1), ether,a mixture of ether and ethyl acetate (9:1) and ethyl acetate. The lastfractions obtained with ether and also the fractions obtained withether-ethyl acetate and ethyl acetate yield, after crystallisation froma mixture of acetone and other, pure A 3:18:20 trioxo llfi hydroxy 21acetoxypregnene or its 11:8-semi-acetal melting at 190192 C.

Example 2 1.12 mg. of the 18:11-lactone of A -3.20-dioxo-11,8:21-di'l1ydroxy-pregnene-18-acid are dissolved in 5 cc. of ethylenedichloride. After the addition of a small crystal of p-toluene sulfonicacid and a drop of pure ethylene glycol, ethylene dichloride is slowlydistilled off. By dropwise addition of new solvent, the volume of thereaction solution is maintained constant. After 4 hours, the content ofthe flask is washed twice with 1 cc. of a 1% sodium carbonate solutioneach time and twice with 1 cc. of water each time and the solution driedand evaporated under vacuum. The residue constitutes the 18: 11- lactoneof the A -3 :20-diethylene-dioxy-115:21-dihydroxypregnene-18-acid, whichis dissolved without purification in 5 cc. of tetrahydrofurane. Aftercooling to -10 to l5 C. there is added dropwise with stirring atetrahydrofurane solution of lithium-aluminium hydride such as iscalculated for the reduction of the lactone to the semiacetal groupwhile taking into account the 21-hydroxyl group. The cooling bath isthen removed and the temperature allowed to rise to about 20 C. Thetetrahydrofurane solution is diluted 'With 50 cc. ether and Washedconsecutively with 0.1-N sulfuric acid and water, dried and evaporatedunder vacuum. The crude A 3:20 diethylene dioxy 115121 dihydroxy 18-oxo-pregnene of the formula:

onion onion CH O I CH0 0/ 1 O/ (L/ HO 0 0 113C 113C 2 ["Q 0 o isdissolved without purification in 50% acetic acid and allowed to standover night at about 20 C. The reaction mixture is extracted twice, eachtime with 5 cc. of chloroform-ether (1:3) and the extracts are Washedtwice, each time with 1 cc. of water, dried over sodium sulfate andevaporated under vacuum. From the crude product which results,aldosterone can be obtained by chromatography.

If an excess of lithium-aluminium hydride is used in the abovereduction, then after the ketal splitting there is obtained the A-3:20-dioxo-l1p:18:21trihydroxy-pregnene of the formula:

CH OH CHzOH Example 3 20.1 mg. of 18:1l-cyclosemiacetal-I8-monoacetateof A 3:18:20 trioxo 11,6121 dihydroxy pregnene are dissolved, in a drynitrogen atmosphere, in 1.5 cc. of a 0.05-rnolar solution of pyridine inanhydrous chloroform, 14.5 mg. of purest p-toluene sulfonic acidchloride added and after placing under vacuum and well mixing, the wholeallowed to stand for 48 hours at room temperature. The reaction mixtureis then diluted with chloroform and ether, washed with0.5-N-hydrochloric acid, 0.1-N-sodium bicarbonate solution and water,dried with sodium sulfate and the solution evaporated. The residue isdissolved in benzene and chromatographed over 1.0 gm. of aluminium oxideby the fractional elution method. The fractions removed with pure etherand ether-ethyl acetate (3:1) crystallize from acetoneether. Theyconstitute mixtures of 21-chloride and 21- tosy-late and are subjectedtogether to the subsequent reaction with sodium iodide.

12.8 mg. of this resulting mixture and 75 mg. of sodium iodide areboiled in 1.0 cc. of acetone for 6 minutes with exclusion of moisture.Thereupon the whole is cooled 13 in ice Water and evaporated undervacuum. The residue is immediately taken up in 0.3 cc. of glacial aceticacid, thesuspension decolorized by rinsing round with a trace of: zincdust and'then again evaporatedunder vacuum. Water is then added andextraction carried out by shaking-:with chloroform-ether (1 :3)..Thecombined, extracts arewashed with 0.3 N-hydrochloric acid, 0.1N-sodiurii bicarbonate. solution and water, dried with sodium sulfateandevaporated. The remaining pale. yellow lacquer is dissolved inbenzene and chromatographed over 0.5 gm. of aluminium oxide. Thefractions removedwith benzene-ether' (1:1), pureether and ether-ethylacetate (9:1), give from acetone-ether pure18:11-cyclosemiacetal-18-monoacetate of M6:18:20-trioxo-1lfi-hydroxyrpregnene.

Example-4.

To a mixture of 8.25 g. of the d:l-A -3-ethylene-dioxy-1113:18-dihydroxy-20-oxo-pregnenedescribed in our application Serial No.480,062, filed concurrently herewith, 4.7. g. offreshlyprepared drysodium methylate and-9.4 g. .of oxalicacid dimethyl ester are added 65cc. of dry benzene. The whole is then stirred for 24 hours in acurrentcfnitrogenat room temperature. After adding ice and-cautiously.acidifying,- the mixture is immediately. ex tracted-by agitation withether. The ethereal-solution'is extracted with'173 cc. of a ,1N-solution of caustic potash, and the. aqueous: alkaline solution isallowed to stand at room temperature for 1 hour. It is then cautiouslyacidifiedwhile cooling withice and. again immediately extracted byagitation with ether. From the residue obtained by. dryingandevaporating in vacuo the ethereal solution there. is obtained .bycrystallisation .d: l-A-3-ethylenedioxy-ILB:18-dihydroxy-aetienoyl-pyruvic acid in the form-ofcolorless crystals.

2.31 g. of the acid so obtained are mixedwith600. cc. of-water whichcontains 25 g. of 'disodium hydrogen phosphate, and thena suspension of1.27 g. of iodine in 120 cc; of-i-ether is slowly. added while stirring.When-the color of the iodine has disappeared; a solution of 1.2 g. ofpotassium hydroxidein- 36 cc. of Water is added, and the whole isstirred in the dark for 24 hours in an atmosphere of nitrogen. To theresidue obtained by separating, drying and evaporating in vacuo theethereal solution are added'8 g. of dry potasisum acetate and 100 cc. ofacetone. After boiling themixture for one hour under reflux, the acetoneisevaporated invacuo and'the residue is extracted several times .withether. From-the-ethereal solutions there is obtained by Washing withwater, drying and evaporating those solutions, crude d:l-A3'-ethylenedioxy '11,8:l8 dihydroxy 20 oxo 21 acetoxy pregnene, which isI obtained in a pure form by chromato raphyv over aluminium oxide.

0.25 g. of the ketal is stirred overnight with a solution of-.t0.05 g.of para-toluene sulfonic acid in 20- cc. of acetone. After dilution witha saturated solution of sodium chloride and extraction by agitation withchloroform; there is obtained from the washed, dried and evaporated'chloroform: solutions dzl-18-hydroxy-corticosterone acetate.

Hydrolysis of the resulting acetate is carried out as follows:

A solution of. 0.3 g. of potassium bicarbonate in cc. of water is addedto 0.25 g. of dzl-18-hydroxy-corticosterone acetate and 40 parts byvolume of methanol in a current of nitrogen, while stirring. Afterallowing the whole to stand for 48 hours at 20 C. in an evacuatedvessel,- the mixture is evaporated in vacuo to one fifth of its originalvolume and extracted by agitation with a mixture of chloroform and ether(1 :3). The residue obtained by'washing with water, drying andevaporating in vacuo the organic solution is chromatographed'over 10parts of aluminium oxide (activity II, neutral), whereby dzl-l8.hydroxyrcorticosterone is obtained.

14 Example 5 0.29, g. of N b'romacetamide' is added (0 0.9 g. of the d:l A5 3 ethylenedioxy- 11,8 18 dihydroxy 20 oxo-21-acetoxy-pregnene.obtained as described in Example 4- and 20 g. of pyridine, Whilestirring. After 4 hours the Whole is poured on to ice and cc. of 2N-hydrochloric acid, and immediately extracted several times withchloroform. In order to separate unreacted starting material, theresidue obtained by drying and evaporating in vacuo the chloroformsolution is chromatographed over aluminium oxide. In this manner thereis obtained dz'l A 3 ethylene dioxy -11 hydroxy 18:20dioxo-21-acetoxy-pregnene.

By using in this example, instead of the ketal, 0.81 g. of'dzl-18-hydroxy-corticosterone acetate, there is obtaineddzl-18-oxo-corticosterone acetate. which exhibits in the infra-redregion characteristic bands at 2.78;. (free hydroxyl group) and at 5.725.82 5.97 1. and 6.16 (double bond region).

0.5 g. of d:l-18-oxo-corticosterone acetate is mixed with-cc; ofmethanol, and then a.solution of 0.6 g. of potassium bicarbonate in 21cc. of water is added in an atmosphere of nitrogen, While stirring. Thereaction solution is allowed to stand for 48 hoursat 20 C. whileevacuated and enclosed. Traces of a precipitate are then filteredpoff,and the solution is extensively evaporated in vacuo. After extractingthe aqueous residue by agitation with a mixture of chloroform and ether(1:3), the organic solution is Washed with Water, dried and evaporated.The resulting d:l-18-oxo-corticosterone is recrystallised from a mixtureofacetone and ether for the purpose of further purification.

The starting materials can be prepared, for example, as follows:

Example 6 A solution of 0.693 gram of 3u-acetoxy llzl7-diketotestane (I)and 0.035 gram of paratoluene sulfonic acid in 14 cc. of glacial aceticacid resisting to chromic acid is. mixed with 2.27 cc. of a 1.06-rnolarsolution of acetoperacid in glacial acetic acid, and themixture ismaintained in the dark for 72 hours at room temperature. A largequantity of Water is then added, the partially precipitated reactionproduct is taken up in methylene chloride, the extract is Washed With an0.5 N-solution of sodium hydrogen carbonate and water, dried with sodiumsulfate and evaporated. The pale yellow residue is dissolved in benzene,and the solution is chromatographed by the fractional elution methodover a column of 21 grams of silica gel prepared with benzene. Aselutriating agents there are used in succession benzene, ether and ethylacetate; At first a small amount of an amorphous substance is dissolvedout with benzene. From the succeeding ether and ethyl acetate elutriatesthere is obtained by crystallisation from a mixture of benzene andhexane pure 3a-acetoxy-13u-hydroxy-13:17secotestane-17-acid-17z13-lactone of the Formula II melting at 178- 180 C.

The same compound can also be obtained by treating 3a-acetoxyor3a-hydroxy-11:20-diketo-pregnane with an aerated and stirred submergedculture of Aspergillus flavus, Penicillium adametzi, Penicillz'umlilacinum or Pencillium chrysogenum, Streptomyces lavendulae, Fusariumsolani or caucasicum, and acylating the reaction product with aceticanhydride in pyridine.

Example 7 chloride is added whilecooling-with ice, the benzeneand theresulting diphenyl are distilled with steam. After cooling the mixture,the precipitated crude product is taken up in ether, the etherealextract is washed with a 2-molar solution of ammonium chloride andwater, dried with sodium sulfate and evaporated.

In order to split oil" water the crude carbinol so obtained is coveredwith 12 cc. of glacial acetic acid and the mixture is boiled for 16hours with the exclusion of moisture. After cooling the mixture, wateris cautiously added and the precipitated reaction product is filteredoff with suction after allowing the mixture to stand for several hours.It is thoroughly washed with Water and dried. By recrystallizing it frommethanol and a mixture of ethyl acetate and isopropyl ether there isobtained pure A -1fi-(3:3-diphenyl-allyl)-7a-acetoxy- 4 keto 2:4bfldimethyl-4am8a5:lOafi-dodecahydrophenanthrene of the Formula IV. In theultra-violet spectrum the compound exhibits a maximum=244 m log 6:452.

Example 8 To a solution of 0.965 gram of A -lfi-(3':3-diphenylallyl)-7ozacetoxy 4-keto-2:4b5-dimethyl-4aa:8afi:10a,8- dodecahydrophenanthrene in11.2 cc. of ethylene chloride and 24.0 cc. of acetic acid of 83.3percent strength there is added at 3 C. while stirring in the course of4 hours a solution of 0.480 gram of chromium trioxide in 4.32 cc. ofacetic acid of 83.3 percent strength. After the mixture has been stirredfor 18 hours while cooling with ice, there are added 7.5 cc. of an 0.1-molar solution of sodium hydrogen sulfite, and the organic solvent isdistilled off in vacuo while replacing the evaporated water. The residueis extracted by agitation with a mixture of ether and methylene chloride(3:1), the extract is first washed with water, 0.5 N-sulfuric acid andwater, and then the acid reaction products are exhaustively extractedwith a 2 N-solution-of sodium carbonate. The combined alkaline extractsare acidified with 2 N-hydrochloric acid and extracted by agitation witha mixture of ether and methylene chloride (3:1). The ether-methylenechloride extract is washed with water and dried with sodium sulfate, andthen mixed with an 0.4l-molar solution of diazo methane in ether untilthe yellow coloration persists, and the extract is evaporated after 20minutes.

The crude ester so obtained is dissolved in 3.0 cc. of anhydrouspyridine and mixed with 2.0 cc. of acetic anhydride. After 16 hours themixture is evaporated in vacuo with therepeated addition of toluene, andthe residue is recrystallised from a small quantity of methanol. Thereis obtained in the form of colorless crystals A 70cacetoxy-4-keto-2:4bfl-dimethyl-4au:8afi:10a}?-dodecahydro-phenanthryl-(1,8)acetic acid methyl ester of the Formula VI,which exhibits in the ultraviolet absorption spectrum a A max 240 mp,log 6:4.20. The acid V can also be obtained directly from the3a-acetoxy- 1317. hydroxy-l3,l7-seco-testane-17-acid-17,13-lactone ofthe Formula II described in Example 6: 20 mg. of lactone II aredissolved in benzene and allowed to stand for 4 hours in a column of 1gram of (neutral) alumina. It is then washed with benzene, ether andchloroform, practically no material being separated. Washing with amixture of methanol and glacial acetic acid then elutriates the A-7a-acet0xy-4-keto-2Abfl-dimethyl-4ao::Safi:lDan-dodecahydrophenanthryl-(1.,B)-acetic acid of the Formula V.

Example 9 1.45.0 gram of MJa-acetoxyi-keto-Zz4bfidimethyl-4aar8af3:10ali-dodecahydrophenanthryl-(1B) acetic acid methyl ester and0890 gram of N-bromo-succinimide are covered with 30 cc. of carbontetrachloride, and the mixture is boiled for 20 minutes under refluxwhile being strongly illuminated. After cooling the mixture in ice waterthe succinimide is filtered off with suction and the pale yellowfiltrate is evaporated in vacuo. .The

16 residue yields by recrystallisation from a mixture of ether andisopropyl ether A 17a-acetoxy-4-keto-2-(bromomethyl) 4bfimethyl-4am:SafizlOafi-dodecahydro-phe nanthry1-(l[3)-acetic acid methylester of the Formula VII.

A solution of 0.883 gram of the crystalline bromide in 40 cc. ofmethanol is mixed with 7.6 cc. of a 2.63 N-methanolic solution ofhydrobromic acid, and the whole is allowed to stand for 22 hours. Afterdilution with a large quantity of water the mixture is extracted byagitation with a mixture of ether and methylene chloride (3:1), theextract is washed with water, 0.5 N-solution of sodium acetate andwater, then dried with sodium sulfate and evaporated. The residue iscrude A 7a oxy-4-keto-2-(bromomethyl)-4b,8-methyl-4aa: SaBAOaBdodecahydro phenanthryl-(1fi)-acetic acid methyl ester of the FormulaVIII. It is advantageously further worked up directly.

The free 7a-hydroxy-compound so obtained is dissolved in 10 cc. ofanhydrous acetone, 0.982 gram of anhydrous potassium acetate is added,and the mixture is boiled for 6 hours with the exclusion of moisture.After cooling the mixture, 20 cc. of water are added and the acetone isdistilled off in vacuo. The precipitated resin is taken up in ether, andthe extract is washed with water; an ice cold 0.1 N-solution of sodiumhydrogen carbonate and water, dried with sodium sulfate and evaporated.For the purpose of purification the residue is dissolved in a mixture ofhexane and benzene (9:1) and chromatographed by the fractional elutionmethod over a column of 25 grams of aluminum oxide prepared with hexane.As elutriating agents there are used a mixture of hexane and benzene(9:1) and (1:1), benzene, a mixture of benzene and ether (9:1) andether. The last portions eluted with benzene and also the fractionssubsequently obtained with benzene-ether (9:1) and ether crystallisefrom a mixture of ether and petroleum ether. By recrystallisation fromthe same pair of solvents there is obtained pure A-7a-hydroxy-4-keto-2-(acetoxy-methyl)- 4hr; methyl-4aa:8a,8:10a/3dodecahydro phenanthryl- (1 ,8)-acetic acid methyl ester of the FormulaIX.

Example 10 1.514 gram of A -7a-hydroxy-4-keto-2-(acetoxy-rnethyl) -4b3-methyl-4ax Safi: IOaB-dodecahydro phenanthryl- (l,8)-acetic acidmethyl ester are added while rinsing with 2.2 cc. of pyridine to asuspension of the chromic acid-pyridine complex prepared by introducing0.440 gram of chromium trioxide into 8.8 cc. of pyridine. The mixture isstirred for 18 hours at room temperature. A large quantity of water isthen added, the mixture is extracted with other by agitation, theextract is washed in succession with water, ice cold 0.5 N-sulfuric acidand water, and then dried with sodium sulfate and evaporated. Theresidue is crystallised from a mixture of ether and isopropylether. Theresulting A 4:7-diketo-2-(acetoxymethyl)4b{3-methyl-4aa: 83cc;IOafl-dodecahydro phenanthryl-(lm-acetic acid methyl ester of theFormula X exhibits, in contradistinction to the starting material, nohydroxyl band at 2.7; in the infra-red absorption spectrum.

0.753 gram of the above diketone is dissolved in 15 cc. of glacialacetic acid, 0.30 cc. of a l-molar solution of hydrogen bromide inglacial acetic acid is added, and 4.2 cc. of a solution of bromine andsodium acetate in glacial acetic acid, which is 0.5-molar with respectto both reagents, is slowly added dropwise while stirring. When thebromine is practically completed consumed, cc. of water are added andthe mixture is extracted by agitation with benzene. The extract iswashed in succession with water, 0.5 N-sodium hydrogen carbonate andwater, dried with sodium sulfate and evaporated in vacuo. The residuewhich slowly crystallizes on standing is crude A 4:7 diketo 2 (acetoxymethyl) 8 bromo- 4b,B methyl 4av.:8afi: l0a,B dodecahydro phenanthryl-1,8)-acetic acid methyl ester of the Formula XI, and is purified byrecrystallisation from methanoland a mixture of ether and isopropylether.

To 0.455 gram of the above bromide in 75 cc. of ethanol of 95 percentstrength is added, while stirring, 0.300 gram of semicarbazide and 0.75cc. of glacial acetic acid. After 16 hours 75 cc. of water arecautiously added, the precepitated semicarbazone is collected on asuction filter after allowing the mixture to stand for one hour, and thefilter residue is washed with water and dried in vacuo. For the purposeof splitting it is covered with 15 cc. of glacial acetic acid and 5.0cc. of an aqueous 0.4 N-solution of pyroracemic acid and stirred for 38hours in an atmosphere of carbon dioxide. After the addition of 20 cc.of water the mixture is extracted by agitation with a mixture of etherand methylene chloride (3:1) the extract is Washed in succession with 2N-solution of sodium hydrogen carbonate, water, 1 N-hydrochloric acidand water, and then dried with sodium sulfate and evaporated. Theresidue is dissolved in a mixture of hexane and benzene andchromatographed over a column of 20 grams of aluminum oxide preparedwith hexane. Elutriation is carried out in succession with a mixture ofhexane and benzene (9:1) and (1:1), benzene, a mixture of benzene andether ('9: 1) and ether, and the fractions dissolved are tested by paperchromatography. The practically unitary benzene elutriates andbenzene-ether elutriates crystallize from methanol. In this manner thereare obtained practically colorless crystals of A-4:7-diketo-2-(acetoxy-rnethyl)--4b,8-methyl 4aazl0afl decahydrophenanthryl (1)3) acetic acid methyl ester of the Formula XII whichexhibits in the ultraviolet absorption spectrum a A max=246 my; log6:4.53.

Example 11 A mixture of 1.498 gram of A -4':7-diketo-2-(acetoxymethyl)4bB methyl 4aaz10afi decahydro phenan thryl-(1[3)-acetic acid methylester, 2.25 cc. of ethylene glycol, 30 cc. of ethylene chloride and0.016 gram of para-toluene sulfonic acid is subjected to slowdistillation for 3 hours during which the moist ethylene chloride whichdistills off is replaced continuously by introducing fresh solvent.After being cooled, the reaction mixture is extracted in succession witha 1 N-solut-ion of potassium carbonate and water, dried with sodiumsulfate and evaporated. The resulting A -4-keto-2-(hydroxy-methyl) 7ethylene dioxy 4b;8 methyl 4am:10a 8- decahydro-phenanthryl-(1p)-aceticacid methyl ester of the Formula XIII is purified by recrystallisationfrom a mixture of ether and petroleum ether.

0.753 gram of the above ketal is dissolved in 20 cc. of anhydroustetrahydrofurane, a solution of 0.950 gram of lithium aluminium hydridein 13.5 cc. of anhydrous tetrahydrofurane is added dropwise, whilestirring, and stirring is continued for 36 hours at room temperature.2.7 cc. of water are then added dropwise while cooling externally withice water, the mixture is freed from precipitated inorganic material,the filter residue being washed with tetrahydrofurane, and the filtrateis evaporated in vacuo. The residue is crude A -4fl-hydroxy 1,3 (2hydroxy ethyl) 2 (hydroxymethyl) 7- ethylene dioxy 4b,6 methyl 4au:10a 3decahydrophenanthrene of the Formula XIV.

The crude triol so obtained is allowed to stand with 3.0 cc. of aceticanhydride in 4.5 cc. of pyridine for 16 hours at room temperature. Thereaction mixture is then evaporated in vacuo with the repeated additionof toluene, the residue is dissolved in benzene, and the solution isfiltered through a short column of active carbon. On evaporation theclear filtrate leaves behind a pale yellow residue. By recrystallisationfrom methanol there is obtained A 4,3 hydroxy 1B (2 acetoxy ethyl) 2-(acetoxy methyl) 7 ethylene dioxy 4b,B methyl-4am:10a/3-decahydro-phenanthrene of the Formula XV.

Example 12 A solution of 1.738 grams of n tp-hydroxy-iys- (2' acetoxyethyl) 2 (acetoxy methyl) 7- ethylene dioxy 4b;8 methyl 4aazl0-alidecahydroph'enanthrene in 13.2 cc. of pyridine is added to 0.440 gram ofchromium trioxide in 8.8 cc. of pyridine, and the mixture is stirred for62 hours at room tem erature. The whole is then diluted with a largeamount of water, extracted by agitation with a mixture of benzene andether (1:2), the extract is filtered through Su er Gel, then Washed withwater, dried With sodium sulfate, ah'd evaporated in vacuo. The residueis dissolved in a mixture of hexane and benzene (3: 1), andchromatographed by the fractional 'elution method over a column of 52grams of aluminium oxide prepared with hexane. Elutriation is carriedout in succession with a mixture of hexaiie and benzene (3:1,), benzene,a mixture of benzen'e'and ether (9:1) and 1:1), ether and a mixture ofether and ethyl acetate (9:1). The fractions are tested by paperchromatography. The practically unitary elutriates which exhibit astrong absorption in the ultra-violet com ta'in A 4 i keto 1p (2"acetoxy ethyl) 2- (acetoxy methyl) 7 ethylene dioxy ib/3 rnethyl-4aor:10addecahydro-phenanthrene of the Formula To 0.865 gram of theabove ketone in 20 cc; of benzene are added 20 cc. of a 1 N-methanolicsolution of potassium hydroxide, and the whole is heated at the boil for2 hours. After cooling the mixture, 10 cc. of

water are added, carbon dioxide is first introduced in order toneutralise the potassium hydroxide, and then the methanol is distilledon? until the final volume of the mixture is small. After the additionof water, the crude product is taken up in a mixture of ether and ethylacetate (1:1), and the extract is washed with water, dried With sodiumsulfate and evaporated in vacuo. The residue consists substantially of A-4-keto-l,8-(2'-hydroxyethyl) 2 (hydroxy methyl) 7 ethylene dioxy-4bp-rnethyl-4aa: IOa/B-decahydro phenanthrene of the Formula XVII, andis then further Worked up immediately.

The crude diol so obtained is dissolved in 4 cc. of

pyridine, 1.41 grams of para-tosylbromide are added while stirring, themixture is initially cooled for one hour to about 10 C., and thenallowed to stand for 72 hours at room temperature while stirringoccasionally. At the end of this period the greater part of the pyridineis evaporated cautiously in vacuo, the crystals are stirred with amixture of methanol and ether (1:1) and the mixture is filtered withsuction. The resulting N-{ [A 4 keto In (2' tosyloxy ethyl) 7 ethylenedioxy- 4bB methyl 4az10afl deeahydrophenan'thryl (2)} methyl}pyridiniumsalt of the Formula XVIII is purified by recrystallisation from amixture of methanol and acetone.

Example 13 10 cc. of an 0.2 N-soluti'on of sodium hydroxide are added toa solution of 1.289 grams of N-{[A 4-keto- 1B (2 tosyloxy ethyl) 7 aethylene dioxy 411 3- inethyl 4aoz10a5 decahydro phenanthryl (2):]-

nitrone of the Formula XIX is crystallized from methanol with the use ofmethylene chloride as solution promoter.

0.635 gram of the nitrone is agitated with 15 'c'c'."of 1 N-hydrochloricacid and 60 cc. of a mixture of ether and benzene (3:1) for one hour inan atmosphere of hydrogen. The organic phase is then washed with water,0.5 N-sodium hydrogen carbonate solution and Water, dried with sodiumsulfate and evaporated in vacuo. I he neutral to litmus. .is then takenup in a mixture of ethyl acetate and benzene '(321), the solution iswashed with 0.01 N-amrnonium carbonate solution and water, dried withsodium sulfate 1 and evaporated. The residue is heated with 25 cc. of.ethylene glycol at 120125 C. until carbon dioxide ceases .to be splitoff, the solution is then cooled and Water is added cautiously. Theprecipitated A -3zl8-bis-ethylene- 19 residue consists of crude A-4:7-diketo-lfi-(2'-to yloxyethyl) 2 formyl 4b/3 methyl 4aa:10a,8decahydrophenanthrene of the Formula XX.

The resulting crude aldehyde is dissolved in 25 cc. of ethylenechloride, 2.5 cc. of ethylene glycol and 0.019 gram of para-toluenesulfonic acid are added, and the mixture is subjected to slowdistillation for 3 hours. The moist ethylene chloride which passes overis continuously replaced by fresh ethylene chloride. After being cooled,the solution is agitated with an 0.5 N-solution of potassium carbonateand water, dried with sodium sulfate and evaporated. The residue isdissolved in a mixture of hexane and benzene 1:1), and for the purposeof purification the solution is chromatographed by the fractionalelution method over a column of 32 grams of aluminium oxide preparedwith hexane. Elutriation is carried out in succession with a mixture ofhexane and benzene (1:1) and (1:3), benzene, and a mixture of benzeneand ether (9:1). The benzene fractions and benzene-ether fractions,which are practically unitary according to tests by paperchromatography, are crystallized from ether, and there is obtained A-4-keto1/8 (2' tosyloxy ethyl) 2 (ethylene dioxy methyl)- 7 ethylenedioxy 4b/8 methyl 4aaz10aB decahydro-phenanthrene of the Formula XXI.

Example 14 2.720 grams of A -4-keto-1,8-(2-tosy1oxy-ethyl)-2-(ethylene-dioxy-methyl)-7-ethylene-dioxy 4b,? methyl-4aa:10aB-decahydro-phenanthrene and 5 cc. of absolute ethanol are addedto a solution of sodium malonic ester,

prepared from 0.130 gram of sodium, 25 cc. of absolute ethanol and 0.883gram of malonic acid diethyl ester, and

the mixture is heated at the boil for 8 hours in an atmosphere ofnitrogen. Water is added to the cooled mixture,

the greater part of the ethanol is distilled off in vacuo, and the crudealkylation product is taken up in a mixture of ether and benzene (1:1).

The solution is washed with water, dried with sodium sulfate andevaporated.

To a solution of the residue in 100 cc. of tertiary butanol there areadded 5 cc. of an 0.1-molar solution of .potassium tertiary butylate intertiary butanol, and the whole is heated on a water bath for 2 hours inan atmosphere of nitrogen. The sclvent is then distilled off in vacuowhile introducing water, the precipitated condensation product is takenup in benzene, the solution is washed with water, dried with sodiumsulfate and evaporated. The residue is dissolved in benzene and thesolution is chromatographed by the fractional elution method over acolumn of 75 grams of aluminium oxide prepared with no absorption in theultra-violet yield when recrystallized from a mixture of ethyl acetateand ether A -3:l8-bisethylene-dioxy-l 1-keto-androstene-l7:17-dicarboxylic acid diethyl ester of the Formula XXIII.

1.065 grams of the dicarboxylic acid ester are saponi- .fied by beingboiled for 2 hours in an atmosphere of nitrogen with 50 cc. of a 4N-methanolic solution of potassium hydroxide. After the addition ofwater, neutral substances which are still present are removed from themixture by agitating it with a mixture of ether and benzene 1:1), andthen 0.1 N-sulfuric acid is added, while cooling with ice and stirring,until the reaction is The precipitated free dicarboxylic acid V 20dioxy-1 l-keto-aetienic acid of the Formula XXIV is filtered off afterallowing the mixture to stand for several hours, and the filter residueis washed with Water and dried in vacuo.

Example 15 A mixture of 0.865 gram of A-3:18bisethy1ene-dioxyll-keto-aetienic acid, 0.756 gram of sodium boronhydride and 40 cc. of absolute methanol is boiled under reflux for 2hours. After cooling the mixture in ice water it is neutralised, whilestirring, by first adding about 10 cc. of 2 N-acetic acid and then wateruntil the separation of the reduction product is complete. By filtering,washing the filter residue with water and drying there is obtained A-3:18-bis-ethylene-dioxy-llfi-hydroxy-aetienic acid of the Formula XXV.

, 2 cc. of trifluoracetic anhydride are added at 0 C. to a solution of0.435 gram of the hydroxy-acid in 5 cc. of pyridine, and the mixture isallowed to stand at room temperature for 4 hours. It is then mixed with5 cc. of dioxane and 2 cc. of water, and the whole is allowed to standfor a further 2 hours. It is then extracted by agitation with a mixtureof ether and chloroform (3:1), the extract solution is washed withwater, dried with sodium sulfate and evaporated in vacuo with therepeated addition of benzene. The residue is crude A-3:l8bis-ethylene-dioxy-l1fi-trifluoracetoxy-aetienic acid of theFormula XXVI.

Example 16 4.597 mg. of aldosterone hydrate are dissolved in 0.7 cc. ofpurest methanol and treated with 0.9 cc. of sodium periodate solution.(This solution is prepared as follows: 1.25 gm. of Na I-I IO aresuspended in about 70 cc. of water and treated with shaking with 2N-sulfuric acid until the reaction'is distinctly acid to litmus,whereupon the whole is made up with water to cc.) After one hour longneedles are formed. After standing for two hours, the methanol isevaporated under vacuum and the residue extracted by shaking first with1 cc. of chloroform, then with 7 cc. of chloroform-ether (1:3) andfinally twice, in each case with 6 cc. of chloroform-ether (1:3). Theorganic solvents are then washed at about 0 C. twice with 0.6 cc. ofsodium carbonate solution and twice with 0.6 cc. of water, then driedover sodium sulfate and evaporated under vacuum. The crystalline neutralportion thus obtained amounts to 3.17 mg. and constitutes the 20:18lactone of the 18:11 cyclo-semiacetal of A-3zl8-dioxo-lIB-hydroxy-aetienic acid of the formula.

After recrystallisation from acetone-ether it is obtained in the form ofcolorless pointed prisms or leaflets of MP. 307-3l3 C. (with browncoloration); [a] +121.2i2 (c.=0.9l54 in chloroform). By concentration ofthe mother liquor, a further quantity of the above lactone can beobtained, which after sublimation under 0.03 mm. pressure and at 192-210C. melts at 307-313 C.

For the isolation of the acid formed in the oxidation, the aqueoussodium carbonate solutions obtained as above are treated at 0 C. with1.4 cc. of 2 N sulfuric acid until the whole is just acid to Congo redand then extraction carried out three times by shaking with 7 cc. ofchloroform-ether (1:3) each time. The product is then in turn washedthree times with 0.5 cc. of water each time, dried over sodiumsulfate'and the solvent 21 evaporated. The crystalline residue amountsto.0.45 mg. and constitutes the crude A -3:18-dioxo-llfi-hydroxyaetienicacid of the formula:

G-OH

CH coon coon This compound yields, on recrystallisation from acetoneether, short colorless prisms which on heating under the microscope areconverted at about 220-240 "C. into transparent leaflets which melt atabout 295-303 C. with brown coloration.

Detection of the formaldehyde formed in the above reaction: The sulfuricacid aqueous solutions obtained above are adjusted with 0.2 cc. of 2 Nsodium carbonate solution to a pH of about -6 and then evaporated tocomplete dryness under 80 mm. pressure and at 60 C. The distillate istreated with 5 mg. of dimedone and 0.5 cc. of glacial acetic acid,heated for one hour to 100 C. and left to stand for 16 hours at about 18C. The separated crystals, after filtration with suction and wash- .ingwith water, amount to 0.66 mg, melt at 192-195" C. and constitute theformaldehyde-dimedone derivative. From the acetic acid aqueous solution,by extraction with :ether, a further quantity can be isolated.

Example 1.7

0.4 mg. of the A -3z18-dioxo-1lp-hydroxy-aetienie acid described inExample 16 are sublimed in a small flask under 0.03 mm. pressure and at210-230 C. The "sublimate is dissolved in 2.5 cc. of chloroform and 7.5cc. of ether and washed with 0.5 cc. of 2 N sodium carbonate solution,0.5 cc. of water, 0.3 cc. of 2 N sodium carbonate solution and thenthree times with 0.3 cc. of water each time. These solutions are furthertreated in a second separating funnel with 4 cc. of chloroform-ether.The chloroform-ether solutions are combined, dried over sodium sulfateand give after evaporation 0.3 mg. of neutral residue which constitutesthe :18-1actone of the 18:11-semiacetal of the A-3zl8-dioxo-llfl-hydroxyaetienic acid, described in Example 16.

Example 18 semiacetal of the A -3:l8-dioxo-llfi-hydroxy aetienic acid,described in Examples .16 and 17, are hydrogenated :in glacial aceticacid solution in the presence of 5 mg. .of prehydrogenated platinumoxide. After, 7 minutes 1.235 cc. (calculated quantity 1.32 cc.) ofhydrogen have By treatment with a mixture of acetic anhydride andpyridine there is obtained the 20: 1-8-lactone of the 18:11-

Y 9.692 mg. of the 20:18-lactone of the 18:1'1-cyclo- 22 cyclosemiacetalof 3fi-acetoxy 11p hydroxy .18 oxoaetianic acid which crystallizes frommethanol in the form of flat needles melting at 218-225 -C.; [a] =--0(c.=0.5246 in chloroform)- v The non-acetylated mixture can be convertedinto the known 3:11-diketo-alloaetianic acid methyl ester in thefollowing manner:

The above hydrogenation product (10.2 mg.) is heated in a mixtureconsisting of 0.03 cc. of hydrazine hydrate and a solution of 12 mg. ofsodium in 0.5 cc. of ethanol in an evacuated bomb tube for 13.5 hours to148 C. After working up there are obtained 3 mg. of neutral portion and7.8 mg. of acid constituents. The latter, after crystallisation fromacetone-ether, yield 3.6 mg. of 18:11-lactone of38:1lfl-dihydroxy-alloaetianic acid- 18-acid of the formula:

0 do on which melts at 270-275 C. (with decomposition). From the motherliquor of the acid there is obtained a further 1.1 mg. of substance ofMP. 252-268 C. (with decomposition). The residue of the acid fraction(3.2 mg.) is amorphous.

The amorphous acid (3.2 mg.) is esterified with diazomethane and themethyl ester thus obtained is oxidized in glacial acetic acid solutionwith 2 mg. of chromium trioxide. After two hours, the oxidation isinterrupted, the reaction solution worked up and the crude productchromatographed over 300 mg. of aluminium oxide. From the crystallizedfractions, after recrystallisation of ether-pentane, crystals areobtained of 'M.P. 205-212 (3., which are identical with known '3:ll-dioxo-alloaetianic methyl ester.

From the neutral portion, the 20:18-lactone of 35:113:l8-trihydroxy-alloaetianic acid of the formula:

The 18: 1 l-lactone of the 3,8: l lfl-dihydroxy-alloaetianicacid-lS-acid (4.7 mg.) obtained according to Example 18, is esterifiedwith diazomethane and the crude product recrystallized fromacetone-ether. The resulting 18:11- lactone of3,8:1lfi-dihydroxy-alloaetianic acid methyl ester-l8-acid of theformula:

0 COOCH:

melts at 232-236" C.; [a] =+81.5i6 (c.=0.362 in chloroform).

3.5 m1. of the above lactone-methyl ester are dissolved pressure.

in 0.4 cc. of purest glacial acetic acid and treated with The above 3B-hydroxy-lactone-methyl ester on acetylation with aceticanhydride-pyridine yields the l8:11-lac tone of 3,8-acetoxy-llfi-hydroxy-alloaetianic acid methyl ester-18-acid. It crystallizes froma mixture of methanol and acetone in the form of rhombic leafletsmelting at 194195 C.; [a] =+64.1 (c.=0.4836 in chloro form).

Example 20 1.267 mg. of aldosterone monoacetate of MP. 190-192" C. aredissolved in 0.12 cc. of glacial acetic acid and treated with 0.01 cc.of 2% chromium trioxideglacial acetic acid solution. After 15 minutesthe chromium trioxide is consumed and a further 0.01 cc. of the samesolution is added. After 3 /2 hours a little methonal is added to thereaction solution and the whole allowed to stand for a further hour andthen evaporate under vacuum at 30 C. The residue is taken up inchloroform-ether (1:3), washed at C. twice with 0.15 cc.

-of N-sodium carbonate solution each time and twice with 0.1 cc. ofwater each time, the chloroform-ether solution dried and evaporatedunder vacuum. The neutral portion is recrystallized twice fromacetone-ether and washed with ether and pentane. The melting point ofthe resulting 18:11-lactone of A-3:20-dioxo-11p-hydroxy-2l-acetoxy-pregnene-18-acid of the formula:

CH OAc which occurs in colorless leaflets (also in part in clusters), isl87-190 C.

After further crystallization from a mixture of chloroform and etherthis compound melts at l98199 C. (fiat needles); [a] =+117.2 (c.=0.6186in chloroform).

For hydrolysis of the acetate group, 6.3 mg. of the above compound aredissolved in 1 cc. of methanol, a solution of 7 mg. of potassiumbicarbonate in 0.24 cc. of water are added and the Whole is allowed tostand at 18 C. for 48 hours. After acidification with dilutehydrochloric acid it is extracted with a mixture of chloroform and ether(1:3), the extract washed first with water,'

then with sodium bicarbonate solution and again with water, dried, andthe solvent evaporated under reduced The neutral portion so obtainedamounts to 3.5 mg. and is tbe l8.11-lactone of A -3.20-dioxo-1 113.21-

'' dihydroxy-pregnene-18-acid. After recrystallization from 24 a mixtureof acetone and ether it forms small grains which melt at 203-218 C.

For oxidatively splitting the ketal group, 5.8 mg. of thislast-mentioned compound are dissolved in 0.6 cc. of methanol, mixed with1.2 cc. of a sodium periodate solution prepared according to Example 16,and allowed to stand at 20 C. for 1% hours. The reaction mixture is thenrendered slightly acid to Congo and extracted with a mixture ofchloroform and ether (1:3). After washing the extract with dilute sodiumcarbonate solution and water, drying, and evaporating the solvent, thereare obtained 3 mg. of a neutral portion. To obtain the acid formed inthe oxidation, the sodium carbonate solutions are acidified andextracted with a mixture of chloroform and ether (1:3). Thechloroform-ether solution, after being washed with water, yields ondrying and evaporation 2.8 mg. of the 18.1l-lactone of the A-3-oxo-1lfihydroxy-aetienic acid-18-acid of the formula which isobtained in the form of fine grains melting at 310-320 C. aftercrystallisation from a mixture of acetone and ether. By methylation bymeans of diazomethane andrecrystallisation from a mixture of acetone andether there is obtained the corresponding methyl ester of melting point219-225 C. (prisms).

' The above mentioned acid melting at 310-220" can be hydrogenatedaccording to the directions given in Example 18. By this means there isobtained the 18:11- lactone of 3B:1l5-dihydroxy-alloaetianicacid-l8-acid of M.P. 270-275 C. (with decomposition), described inExample 18.

1.7 mg. of the above-described 18:11-lactone of the A-3-oxo-1lfi-hydroxy-aetienic acid methyl ester-18-acid of melting point2l9-225 C. are dissolved in 1.5 cc. of glacial acetic acid andhydrogenated at 22 C. in the presence of 1.5 mg. of platinum oxide. Whenthe hydrogenation ceases the catalyst is removed by filtration, thefiltrateevaporated under reduced pressure and the residue taken up in amixture of chloroform and ether (1:3). The chloroform-ether solution isthen washed twice with water, dried and evaporated in vacuo. The cruderesidue amounts to 1.7 mg. and melts at 210-230 C. afterrecrystallisation from a mixture of acetone and ether. On furtherrecrystallizing, the melting point rises to 232236 C. This compound isidentical with the 18,11-lactone of 313,1lfl-dihydroxy-alloaetianic acidmethyl ester-18-acid described in Example 19.

Example 21 7 mg. of aldosterone hydrate are dissolved in 0.7 cc. ofmethanol and treated with 1.28 cc. of sodium periodate solution(prepared according to Example 16). After only a few minutes, theseparation of crystal needles commences. After 2 hours the methanol isremoved under vacuum at 20 C. Then at 0 C., with 2 N-H SO a justdistinctly acid reaction to Congo red is established and the wholeextracted by shaking three times, in each case with 10 cc. ofchloroform-ether (1:3). (For the Working up of the aqueous phase seebelow.)

The three extracts are washed in turn by means of the followingsolutions: 1 cc. of Water, 1 cc. of 0.2 N-sodium carbonate, 0.5 cc. ofwater, l'cc. of sodium carbonate solution and twice with 0.5 cc. ofwater each time. The solutions, dried over sodium sulfate, give onevaporation 6.7 mg. of neutral constituents. The combined water andsodium carbonate extracts are acidified with sulfuric acid and extractedby shaking-with chloroform- COOH 25 (.113). (For the working up of theacid aqueous phase see below.) The extracts washed with a. little water.and dried over sodium sulfate give on'evaporation 0.4 mg. of 11-3:18-dioxo-11,8-hydroxy-aetienic acid (see Example .16,)

The 6.7 mg. of neutral constituents give from acetoneether 4.5 mg. ofthe 20: 1'8-lactone, describedin Example .16, of the18::11-cyclosemiacetalof A -3z18-dioxo-llfihydroxy-aetienic acid of M.P.309-315 'C. (with decomposition). The mother liquors, after distillationin 'a small flask or after chromatography-over alumina, give a furtherquantity of the same compound. a Detection of the formaldehyde formed inthc oxidation: The two acid aqueous phases, after extraction withchloroform-ether, are combined, brought to pI-I'=5 with sodium carbonatesolution and distilled to dryness under 80min. pressure and at 60 C.bath temperature. The distillate subjected to "repeated distillation.The distillate thus purfied is treated with 5 mg. of dimedone and 0.3cc. of glacial acetic acid, heated :for 1 hour to 100 C. and allowed tostand for 12 hours at 0 C. 0.66 ,mg. of needles of M.P. 192-194" C.crystallize. The moth'er liquor is extracted with ether. The solution,washed with a little water and dried over sodium sulfate, is evaporatedand the residue chromatographed over 0.2 gm. -of Mg-silicate-kieselguhr(Celite No. 545) (2:1). The fractions eluted with benzene (0.8 mg.) givefrom ether-pentane a further small quantity offormaldehydedimedonecompound (M.P.: about 180 C.). The .fractionselutable with benzene-ether and pure ether give freedimedone, M.P..130-135 C. I .The' combined crystals of the formaldehyde compound aresublimed in a small flask under 0.02 .mm. pressure and at 105-115 C. Thesublimate gives from etherp'cntane colorless crystals of M.P. 192-194 C.A known sample of formaldehyde-dimedone and a mixture of the twosubstances both melt at the same temperature. The infra-red spectra arealso the same.

. Example 22 "69 mg. of crude aldosterone-hydrate are dissolved in 6 cc.of methanol and allowed to stand for 11/2 hours with 12 cc. of sodiumperiodate solution. Working up by a method analogous to the directionsof Examples 16 Iand 21 gives 21 mg. of the A-3:l8-dioxo-llfi-hydroxyaetienic .acid described in Example 16 and 37.8mg. of a neutral fraction. The latter gives from acetone-ether 4.7 Ir1g.,o'f crude 20:18-lactone of the 18:1l-cycloscmiacetal .of 21-3:l8-dioxo-l1B-hydroxy-aetienic acid of M.P. 285-305" C. (withdecomposition).

In an analogous manner there are obtainedfrom 195 mg. of crudealdosterone-hydrate '95 mg. of the A -3:18- dioxo-ll ti-hydroxy-aetienicacid described in Example 16 and 76 mg. of a neutral fraction. Thelatter yields from acetone-ether 15 mg. of the above described lactoneof M.P. 303-312 'C. (with decomposition).

The mother liquors from the lactone obtained in these two cases aredistilled in a small flask under 0.02 mm. pressure. The fractionspassing over up to 195 C. bath temperature are discarded. The distillateobtained from 195-230 C. gives from acetone-ether a further quantity ofabout 0.7 mg. of crystalline'lactone of M.P. 275-295 C. Chromatographyof the remaining mother liquors over alumina (alkali free) includingelution with petroleum ether-benzene and pure benzene, gives fractionscontaining only further traces of the crystalline lactone.

The not quite pure fractions of the crystalline lactone are 'subli'medin a small flask under 0.02 mm. pressure and at 210-230 C. bathtemperature. The sublimate gives-from acetone-ether directly purecrystals of M.P.

307-311" C. (with decomposition).

Example 23 I 1. 1.9 mg. of aldosterone-hydrate are dissolved in 0.05 cc.of purest glacial acetic acid and treated with 0.01165 26 cc. of 2%chromium trioxide-glacial acetic acid solution (equals 0.33 mg. of CrO Ayellow brown precipitate (-CrO -complex) is immediately produced whichdissolves only gradually. The chromium-trioxide becomes spent afterabout 2 minutes and then consecutively four equal portions of chromiumtrioxide solution are further added (total therefore 1.65 mg. CrO =5.0equivalents). The first of these portions becomes spent after 5 minutes,the second after 10 minutes and the third after 45 minutes, whereas thefourth is not completely consumed after a further 1% hours. The whole isthen evaporated under vacuum at 30 C., treated with water and 3 drops/of2 N-sulfuric acid and extracted with chloroform-ether 1:3). Theseparation according to the directions of Examples 16 and 21 yields atrace of acid and 1.8 mg. of neutral constituents, the latter givingfrom acetoneether 0.9 'mg. of colorless granules of M.P. 298- 310 C.This compound can likewise be sublimed without decomposition under 0.01mm. pressure and at 200-230" C. By a mixed melting point test and apaper chromatogram, it is proved to be identical with the lactonedescribed .in Examples 16, 21, and 22. The polarity in the B -system ofBush is about the same as or a little higher than adrenosterone. In theformamidecyclohexane-benzene- (1:1) system at C. R =0.34 (adrenosterone'R =0.59). Example 24 7.3 mg. of 2l-monoacetyl-aldosterone of M.P. 198--199" C. are dissolved in 0.7 cc. of purest glacial acetic acid andtreated with 0.055 cc. of 2% chromium trio'x'id'e 'solution (=1.1 mg.CrO which becomes spent after 25 minutes at 20 C. Another 0.030 cc. ofthe same solution (:06 mg. CrO is then added which after a furtherminutes also becomes spent. A third addition of 0.030 cc. (:06 mg.) isunconsumed after a further 1% hours. The whole is treated with 1 drop ofmethanol and allowed to stand for a further 2 hours. Working upaccording to the directions of Example 21 gives no acid constituents but7.3 mg. of a neutral crude product. From acetone-ether 6.4 mg. ofcrystals of M.P. 196-199 ;C., and 0.1 mg. of M.P. 194-197" C. areobtained. Recrystallization from acetone-ether gives long, narrowhexagonal leaflets of M.P. 198-200 C. and repeated recrystallizationfrom methanol-ether gives 5.5 mg. of hexagonal (to some extent havingrhombic bounding surfaces) leaflets of M.P. 193-194 C.

('c.='0.6186 in chloroform). Reduction test: positive; ultravioletabsorption: positive. In a paper chromatogram (propylene glycol-toluene)this compound migrates 'in the same way as1l-dehydro-Zl-acetyI-corticosterone. This compound constitutes the18:11-1actone, described in Example 20, of the h -3z20-dioxo-lle-hydroxy2'l-acetoxy-pregnene-l8-acid.

Example 25 20.1 mg. of the 20:18-lact0ne, described in Example 16, ofthe 18:11-cyclosemiacetal of A -3:l8-dioxo-1'1B- hydroxy-aetienic acidof M.P. 304-310 C. (with decomposition) are hydrogenated in 2 cc. ofglacial acetic acid with 1.75 mg. of previously hydrogenated PtO Theabsorption of hydrogen is complete after minutes (3.3 cc. H absorbed at22.4 C. under 731 mm. pressure corresponding to 2.14 mol). The crudehydrogenation product is allowed to stand for 16 hours at 20 C. in 0.2cc. of acetic anhydride and 0.3 cc. of absolute pyridine. The whole isthen evaporated under vacuum at 40 C., taken up "in chloroform-ether(1:3), Washed, in each case using 0.3 'cc., with dilute hydrochloricacid, water, potassium bicarbonate solution and water (twice), driedover sodium sulfate and evaporated. The residue (20 mg.) gives from alittle acetone-ether (about 1:10) '8 mg. of fine needles of 199-220 C.Purification is best effected by dissolving in acetone-ether (1:1),adding methanol and concentrating to a considerable extent, as a resultof which the remaining solvent is practically pure methanol. 4.2 mg. offlat needles result, which on heating are converted at about 219 C. intothin long needles of M.P. 222-226 C. This compound is the 20:18-lactone,described in Example 18, of the 18:11-cyclosemiacetal of 3fl-acetoxy-1l-fi-hydroxy-l8-oxo-aetianic acid.

Example 26 9.2 mg. of the 20:18-lactone, described in Examples 18 and25, of the 18:11-cyclosemiacetal of3fi-acetoxy-11fihydroxy-lS-oxo-aetianic acid of M.P. 218-225" C. areheated in a small bomb tube with 0.12 cc. of hydrazine hydrate for 1 /2hours to 100 C. while the tube is open. Then a solution of 11 mg. ofsodium in 0.5 cc. of ethanol is added and the tube is sealed while under80 mm. pressure and heated for 14 hours to 151 C. After cooling, thealcohol is removed under vacuum and the residue treated with a littlewater and extracted by shaking three times, in each case with 10 cc. ofcholoroform-ether. The extracts washed with water and dried over sodiumsulfate leave on evaporation 0.8 mg. of residue (neutral fraction).

The aqueous phase and the wash waters, at C., are brought to pH:-1 withhydrochloric acid and again extracted three times by shaking withchloroform-ether (1:3). These three extracts are washed consecutively incounter current, using 0.3 cc. in each case, with the following liquids:Water, 2 N-sodium carbonate solution, water, 2N-sodium carbonatesolution, water, water. Drying over sodium sulfate and evaporation give1.4 mg.

of a residue of the 20:18-lactone, described in Example 18, of3B:1113:18-trihydroxy-alloaetianic acid, which compound partiallycrystallises.

The sodium carbonate extracts and wash waters are combined, treated at 0C. with hydrochloric acid until the reaction is acid to Congo red andagain extracted with chloroform-ether. The extracts, washed with waterand dried over sodium sulfate, give on evaporation 6.7 mg. of residue(acids).

Separation of the acids: The 6.7 mg. of acid mixture gives fromacetone-ether in two fractions 4.5 mg. of crystals of melting point251-268 C. and as a third fraction 0.5 mg. of melting point 235-265 C.As in this case purification is a difficult matter, the combinedcrystals (5 mg.) and the residue from the mother liquors (1.7 mg.) areesterified with diazomethane.

The 5 mg. of crystalline acid give 5.1 mg. of crude methyl ester.Recrystallization from acetone-ether yields a mixture of coarse prismsand fine needles. The latter can be washed out with ether-pentane. Bycareful fractionation and repeated recrystallization 1.7 mg. areobtained of coarse prisms of M.P. 232237 C. of the 18:11-lactone,described in Example 19, of the 35:115-

dihydroxyalloaetianic acid methyl ester-18-acid. The remaining motherliquors (3.4 mg.) are combined with the methyl ester mixture (1.8 mg.)from the 1.7 mg. of residue from the mother liquors of the crystallineacid. This combined material (5.2 mg.) serves for acetylation (seebelow).

mg. of methyl ester mother liquors are acetylated as described above andthe acetate mixture (5.6 mg.) chromatographed over 500 mg. of alumina.

The fractions 1-12 (eluted with petroleum ether-benzene rising to 50%benzene content) give only alittle residue and at the most traces ofcrystals.

Fraction 15 (eluted with pure benzene) gives from ether-pentane fewflattened, doubly pointed needles of a secondary product of M.P. 198-201C. The mother liquor is combined with fractions 13-14 and gives from alittle ether with pentane few thick parallelepipeds of M.P. 182-185 C.

Fractions 16-18 (eluted With pure benzene and benzene-ether (:5)) givefrom ether-pentane 1.018 mg.

'of pure 18:11-lactone of 3B-acetoxy-11,3-hydroxy-allo- 19.1 mg. of aresidue from the mother liquor of the pure 20:18-lactone of the18:11-cyclosemiacetal of 3B- acetoxy-l1,8-hydroxy-18-oxo-aetianic acidare heated for 2 hours to C. with 0.3 cc. of hydrazine hydrate and afterthe addition of 25 mg. of sodium in 1 cc. of ethanol, heated for 17hours to 151 C. Working up as in EX- ample 26 gives 4.5 mg. of a neutralfraction, 4.4 mg. of the 20:18-lactone, described in Example 18, of35111518- trihydroxy-alloaetianic acid and 8.5 mg. of acids. The 8.5 mg.of acids are methylated (9.3 mg.) and acetylated (10.2 mg). Theresulting material is chromatographed (40 fractions) over 1 gm. ofalumina.

From fractions 13-24 2.7 mg. of crude 18:1l-lactone of3fi-acetoxy-1IB-hydroxy-alloaetianic acid methyl ester- 18-acid of M.P.l85-193 C. are obtained.

Fractions 25-27 give a little impure 18:11-lactone of Sfi-acetoxy-llfl-hydroxy-alloaetianic acid methyl ester-18- acid of M.P. l82192 C.

Fractions 28-38 give 1.6 mg. of pure 3 3-acetoxy-11Bhydroxy-alloaetianic acid methyl ester of M.P. 192-199 C.

Example 28 5.8 mg. of the 20:18-lactone, described in Examples 16, 26and 27, of 35:113:18-trihydroxy-alloaetianic acid are dehydrogenatedwith chromium trioxide in glacial acetic acid, until at 20 C. no morechromium trioxide is consumed. The customary working up gives 4.4 mg. ofa neutral crude product which is chromatographed over 0.3 gm. ofalumina. The fractions (0.4 mg.) eluted with ether-methanol (8-50%methanol content) crystallize. Double recrystallization fromacetone-ether yields colorless fine needles combined in clusters, ofM.P. 275 290 (with conversion into coarse needles) consisting of the 20:18-lactone of 3:11-dioxo-18-hydroxyalloaetianic acid of the formula:

0 oinmo The above examples clearly demonstrate that according to theprocess of the invention either aldosterone or its functionalderivatives or intermediates which are convertible into these compoundsare obtained. Thus the compounds of the invention can be used asmedicaments having the activity of hormones of the adrenal glands or asintermediates for the production of such medicaments. 7

29 What is claimed is: 1. A process which comprises reacting the A-3:18-bisethylenedioxy-l1,8trifluoracetoxy-etienic acid chloride withdiazomethane.

2. A process which comprises reacting A -3:l8-bis- 5 wherein Rrepresents a member selected from the group consisting of a freehydroxyl group and a hydroxyl group esterified with a lower aliphaticcarboxylic acid, R an aldehyde group acetalized with a lower alkanedioland Hal is a halogen atom, and a 3-1ower alkylene ketal of and treatingthe resulting diazo ketone with a member selected from the groupconsisting of Water, an aqueous inorganic acid, an aqueous organiccarboxylic acid and an anhydrous organic carboxylic acid.

6. A member selected from the group consisting of a compound of theformula:

and a 3-lower alkylene ketal thereof, wherein R represents a member ofthe group consisting of a free hydroxyl group and a hydroxyl groupesterified with a lower aliphatic carboxylic acid and R is a member ofthe group consisting of an aldehyde group and an aldehyde groupacetalized with a lower alkanediol.

7. A process of claim 5 wherein the aldehyde group acetalized with alower alkanediol in the resulting diazo ketone is hydrolyzed to the freealdehyde.

8. A compound of claim 6 wherein R is a hydroxyl group esterified withtrifluoroacetic acid.

References Cited in the file of this patent Helv. Chem. Acta., vol. 37(1954), June 15, 1954,

such halide, with an excess of a diazomethane solution 30 pages1163-1223 UNITED STATES PATENT OFFICE Certificate of Correction PatentNo. 2,904,545 September 15, 1959 Tadeus Reichstein et a1.

It is hereby certified that error appears in the printed specificationof the above numbered patent requiring correction and that the saidLetters Patent should read as corrected below.

Column 5, line 28, for pregnene read pregnane; line 52, for acetianicread aetianic; line 53, for 17w read -17a-; columns 7 and 8, FormulaVIII should appear as shown below instead of as in the patent:

columns 9 and 10, Formula XVI should appear as shown below instead of asin the patent:

same columns and 10, Formula XXVI should appear as shown below insteadof as in the patent:

OOOH

column 11, line 66, for 11: 8-semiread -11:18-semi--; column 14, line12, for --3-ethylene-dioxyread ---3-ethylenedioxycolumn 16, line 2, forA l'fw read A -7aline 15, for -oxyread -hydroxycolumn 19, line 56, afterultraviolet insert a comma; column 20, line 47, for 18:11 cyclo read 18:11-oyclocolumn 21, line 62, for acetienic acid read aetianic acids;column 22, lines 63 to 72, inclusive, the. formula should appear asshown below instead of as in the patent:

COUCH:

same column 22, line 75, for 3.5 m1. read 3.5 mg.; column 23, line 74,for 18.11: lactone of A*-3.20-dioxo-11B.21- read. -l8,11-1aotone of A-3,20-di0xo-11,B,21-; column 24, line 17, for 18.1l-1act0ne read18,11-1act0ne; line 34, for 310220 read 310320.

Signed and sealed this 26th day of July 1960.

Atfiesfi:

KARL H. AXLINE, Attesting Oflicer.

A ROBERT c. WATSON, C'ommissz'oner of Patents.

6. A MEMBER SELECTED FROM THE GROUP CONSISTING OF A COMPOUND OF THEFORMULA: